Impact of BDNF -196 G>A and BDNF -270 C>T polymorphisms on stroke rehabilitation outcome: sex and age differences.

BACKGROUND: Genetic factors, including gene polymorphisms, are promising in determining stroke rehabilitation outcome. Brain-derived neurotrophic factor (BDNF) is one of the most attractive because of its role in neuroplasticity and brain repair. OBJECTIVE: The aim of present study was to assess the role of BDNF -196 G≯A (val66met) and -270 C≯T on clinical parameters and functional outcome in patients with ischemic and hemorrhagic stroke. Additional analyses according to sex and age (≤55 and ≯55 years) were performed. METHODS: Three hundred thirty-eight patients (287 with ischemic and 51 with hemorrhagic stroke) were evaluated in terms of neurological deficit (National Institute of Heath Stroke Scale [NIHSS]), activities of daily living (Barthel Index [BI]), and everyday functionality (Rankin score [RS]) before and after rehabilitation. BDNF polymorphism genotyping was performed by polymerase chain reaction restriction fragment length polymorphism analysis. RESULTS: In multivariative analysis, unfavorable outcome of stroke rehabilitation (RS ≥2) was associated with independent factors: ischemic stroke (odds ratio [OR], 2.59; 95% CI, 1.03-6.47), female gender (OR, 2.80; 95% CI, 1.39-5.64), depression (OR, 4.24; 95% CI, 1.45-12.35), falls (OR, 2.61; 95% CI, 1.16-5.87), and BDNF -196 GG polymorphism (OR, 2.18; 95% CI, 1.09-4.35). The differences of functional parameters measured with BI and RS on admission and at discharge are apparent only for comparisons between patients ≤55 and ≯55 years old carrying BDNF -196 GA+AA genotypes but not in those carrying -196 GG genotype; the differences were evident in women but not in men. CONCLUSIONS: BDNF -196 G≯A polymorphism might affect functional outcome of stroke rehabilitation, but this hypothesis needs further verification.

Neurochemical and behavioral characteristics of toxic milk mice: an animal model of Wilson's disease.

Toxic milk mice have an inherited defect of copper metabolism. Hepatic phenotype of the toxic milk mice is similar to clinical findings in humans suffering from Wilson's disease (WND). In the present study, neurotransmitter system and locomotor performance in toxic milk mice was examined to verify the feasibility of this animal model for studying neuropathology of WND. Mice aged 2 and 12 months were used in the experiment. The mice were tested according to rotarod and footprint protocols. Monoamine content in brain structures was measured by high performance liquid chromatography. In order to detect neuronal loss, expression of enzymes specific for dopaminergic [tyrosine hydroxylase (TH)], noradrenergic (dopamine beta-hydroxylase) and serotoninergic [tryptophan hydroxylase (TPH)] neurons was analyzed by Western blot. The 12-month-old toxic milk mice demonstrated impaired locomotor performance in behavioral tests. Motor deficits were accompanied by increased copper and serotonin content in different brain regions and slight decrease in dopamine concentration in the striatum. The expression of TH, dopamine beta-hydroxylase and TPH in the various brain structures did not differ between toxic milk mice and control animals. Despite differences in brain pathology between humans and rodents, further exploration of neuronal injury in toxic milk mice is warranted to broaden the understanding of neuropathology in WND.

New single nucleotide polymorphisms associated with differences in platelets reactivity in patients with type 2 diabetes treated with acetylsalicylic acid: genome-wide association approach and pooled DNA strategy.

The objective of this study was to use genome-wide association approach and pooled DNA strategy to search for new genomic loci associated with inter-individual differences in platelet reactivity in the diabetic patients during acetylsalicylic acid (ASA) treatment. Study cohort consisted of 297 diabetic patients who had been taking ASA (75 mg daily) for at least 3 months. We tested association of single nucleotide polymorphisms (SNPs) genotyped using high density microarray platform with several platelet reactivity assays, followed by individual genotyping of most significant SNPs identified in the microarray genomic scan. The highest statistical significance (p value of 0.0001-0.008 in individual genotyping) was observed for SNP located within the regulatory G-protein signaling (RGS) 7 gene (rs2502448) using recessive genetic model. The diabetic patients on ASA treatment and homozygotes for its minor allele were characterized by increased odds ratio of at 3.45 (confidence interval: 1.82-6.53) for high on ASA platelet reactivity (i.e. impaired ASA response) when compared with homozygotes for wild-type allele. The genome-wide approach might provide an opportunity to identify novel candidate genes and pathways related to platelet activation in diabetic patients.

Effect of common single-nucleotide polymorphisms in acetylsalicylic acid metabolic pathway genes on platelet reactivity in patients with diabetes.

BACKGROUND: Platelet reactivity in patients on acetylsalicylic acid (ASA) therapy can be influenced by physiological or pathological conditions affecting ASA pharmacokinetics or pharmacodynamics. The mechanism of such variability in the therapeutic response to ASA, particularly in diabetic patients, is poorly understood. The rate of elimination of ASA and its metabolite, salicylic acid (SA), is likely a major factor determining drug efficacy. The objective of this study was to investigate the effect of genetic polymorphisms in the selected candidate genes within the ASA metabolic pathway on the platelet reactivity and concentration of ASA and thromboxane A(2) (TxA(2)) metabolites in a population of patients with type 2 diabetes mellitus (T2DM). MATERIAL AND METHODS: The study cohort consisted of 287 Caucasians with T2DM who had been taking ASA tablets at the dose of 75 mg per day for at least 3 months. Platelet reactivity analyses were performed using VerifyNow Aspirin and PFA-100 assays. The measured ASA metabolite included salicylic acid (ASA), and TxA(2) metabolites included serum TxB(2) and urinary 11-dh-TxB(2). Genotyping for the selected 18 single-nucleotide polymorphisms (SNPs) within 5 genes of the ASA metabolic pathway was performed using a Sequenom iPLEX platform. RESULTS: No statistically significant association was observed between the investigated SNPs genotypes, platelet reactivity, and measured metabolites in the investigated cohort of patients. CONCLUSIONS: The results of our study failed to confirm that the selected variants in the genes within the ASA metabolic pathway might contribute to platelet reactivity in a diabetic population treated with ASA.

Influence of BDNF polymorphisms on Wilson's disease susceptibility and clinical course.

Susceptibility to Wilson's disease (WD) and its clinical manifestations are thought to be affected by genetic factors, including polymorphisms. The role of brain-derived neurotrophic factor (BDNF) in the pathogenesis of neurodegenerative diseases is now widely discussed. The aim of the present study was to evaluate the frequency of the BDNF Val66Met (G-196A) and C-270T polymorphisms in WD patients and in healthy controls, and to determine the role of these polymorphisms in the clinical characteristics of WD. We found that the BDNF Val/Val (-196 G/G) and -270 C/T genotypes occurred more frequently in WD patients than in healthy controls (66 % versus 45.5 %, p = 0.0001, and 14 % versus 6 %, p = 0.018, respectively). Similarly, symptomatic patients carried the BDNF Val/Val genotype more often than presymptomatic patients (75 % versus 53 %, p = 0.0097). No association was detected between any of the determined polymorphisms and the dominant form of the disease or the age of onset for WD.

Long-term effect of high doses glucocorticosteroids on mRNA expression for IL-6 and IL-8 in relapsed multiple sclerosis patients.

Glucocorticosteroids (GS) are standard treatment of multiple sclerosis (MS) relapse, but no superiority of any commonly used doses is known. The aim of present study was to evaluate mRNA expression for two cytokines: IL-6 and IL-8. Ethylenediaminetetraacetic acid blood samples from 35 MS relapse patients were obtained before therapy, after 7, 14 days, and 3 months from treatment start (500 versus 1000 mg for 5 days). Significant neurological improvement measured with EDSS was independent to GS dose. Changes of mRNA cytokines expression were more evident in higher dose group but for IL-6 mainly in females.

Care for patients after stroke. Results of a two-year prospective observational study from Mazowieckie province in Poland.

BACKGROUND AND PURPOSE: Little has been published about different elements of the health services in long-term follow-up in the countries of Central and Eastern Europe. The aim of this study was to explore the health services for stroke patients in Poland. MATERIAL AND METHODS: Patients from 3 centres representing different levels of stroke care organization from Mazowieckie province were included. Data on first-ever stroke patients with "onset-to-door" time no longer than 7 days, consecutively admitted to participating centres between March 1 and June 30, 2002 were collected prospectively. Patients were assessed on admission, on discharge and 3, 6, 12, 18, and 24 months after discharge. Type of care, rehabilitation, readmissions, consultations and diagnostic procedures were evaluated. RESULTS: One hundred and sixty-four patients with first-ever stroke were included. Twenty-one patients died during hospitalization, and 36 during the two-year follow-up. Most patients were discharged home, under family care. The total rate of readmission decreased over time, from 58% to 11%, and so did the rate of rehabilitation, from 41.5% to 15%. All patients had been seen by their general practitioners and neurologists. CONCLUSIONS: Post-stroke care is provided mostly by family members. Access to rehabilitation is limited and decreases over time. This study could help the authorities in healthcare budget allocation in Poland.

BDNF A196G and C270T gene polymorphisms and susceptibility to multiple sclerosis in the Polish population. Gender differences.

Two single nucleotide polymorphisms of the human BDNF gene, G196A and C270T, and their impact on the susceptibility to MS and disease progression in the Polish population were investigated. Increased risk of the disease was found for 196G/G carriers (OR -- 1.63, p=0.01) (only females) and 270C/T carriers (OR -- 7.76, p<0.001) (both males and females). The first signs of the disease appeared earlier in 196G/G than 196G/A patients (p=0.01), but it was limited to males. Our results show that C270T and G196A BDNF polymorphisms may affect susceptibility to and onset of MS, but further verification is needed, with special attention to gender differences.

Influence of age and gender on cytokine expression in a murine model of Parkinson's disease.

OBJECTIVE: The neuroinflammatory reaction has been linked with Parkinson's disease. One of the hypotheses to explain the significance of age and gender (male predominance) effects on neurodegeneration in Parkinson's disease may result from a link between these risk factors and the inflammatory processes. Here, we investigated the expression of inflammatory mediators in relation to 1-methyl-4-phenyl-1,2,3,6-tetrahydropiridine (MPTP)-induced neurodegenerative processes in nigrostriatal pathway in young and aged male and female mice. METHODS AND RESULTS: We simultaneously assessed striatal tyrosine hydroxylase (TH) protein concentrations (Western blotting) and cytokine (TNFalpha, IFNgamma, IL-1beta, IL-6 and TGFbeta(1)) mRNA levels (RT-PCR) in young and aged (2- and 12-month-old) C57BL/6 male and female mice after 6 h, 1, 3, 7, 14, 21 days after MPTP intoxication. Western blotting analysis showed that at the early time points, males showed a greater reduction in striatal TH versus females. Additionally, in contrast to the aged mice, in young males and females the TH concentration gradually increased between the 7th and the 21st day after intoxication. The increases in TNFalpha, IL-1beta and IFNgamma after intoxication were faster in both young and aged males than females. In males (both ages), we observed an increase in TGFbeta(1) at the early time points. In contrast, in females (both ages) TGFbeta(1) was elevated at later time points. MPTP caused an increase in IL-6 in males and females, but this increase was significantly higher in females. CONCLUSIONS: A gender and age skewing of the cytokine gene expression in the striatum after intoxication may be related to the greater susceptibility in males as well as older animals to the detrimental effects of MPTP.

Dopamine, serotonin and noradrenaline changes in the striatum of C57BL mice following myelin oligodendrocyte glycoprotein (MOG) 35-55 and complete Freund adjuvant (CFA) administration.

Many data suggest involvement of inflammation in neurodegeneration. However, the exact mechanisms of this cooperation are poorly understood. We have previously shown that induction of inflammatory reaction, both before and after injury of the striatum, affects regeneration of dopaminergic neurons. In the present research we studied the role of inflammatory reaction in non-injured striatum. We used myelin oligodendrocyte glycoprotein (MOG) 35-55 in complete Freund's adjuvant (CFA) to elicit experimental autoimmune encephalomyelitis (EAE) mice model. As determined by HPLC, striatal dopamine (DA) and serotonin levels in mice treated with either MOG 35-55 in CFA or CFA alone were significantly higher compared to vehicle-treated controls on 13th day after induction. The ratio of homovanilic acid/dopamine (HVA/DA) and 3, 4 dihydroxyphenylacetic acid/dopamine (DOPAC/DA) were significantly lower in the MOG and CFA groups on 13th day, indicating decreased DA metabolism. Noradrenaline (NA) concentration did not differ between groups. Moreover, the striatal mRNA IL-1beta and TNF-alpha levels were elevated during induction phase of EAE in both groups, as determined by RT-PCR. Our data indicate regulatory connection between dopaminergic and immune systems.

Mechanism of action of three newly registered drugs for multiple sclerosis treatment.

Multiple sclerosis (MS) is a disease of suspected autoimmune origin leading to neurodegeneration. The disease pathomechanism is considered to be primarily based on neuroinflammation directed against myelin antigens caused by autoreactive T cells. MS etiology remains still unknown, which makes it difficult to create an efficient therapy, therefore, MS treatment targets mechanisms involved in disease pathology. In this review, we present the mechanism of action of three newly registered drugs for MS. Dimethyl fumarate (DMF) is an agent presenting a broad spectrum of action. Its main activity is based on activating the nuclear factor E2 dependent pathway leading to antioxidant enzyme synthesis. DMF in general suppresses the pro-inflammatory immune activity and exerts a neuroprotective action. Teriflunomide is a more focused drug, acting as an inhibitor of pyrimidines synthesis, important for rapidly dividing cells such as activated lymphocytes. Similarly, alemtuzumab, an anti-CD52 antibody, causes depletion of mainly lymphocytes. Since in MS pathology, T and B cells are involved, this mode of action is promising.

High dose of intravenously given glucocorticosteroids decrease IL-8 production by monocytes in multiple sclerosis patients treated during relapse.

The aim of our study was to determine whether high doses of intravenous methylprednisolone have significant impact on immune parameters during the multiple sclerosis (MS) exacerbations. Peripheral blood of 32 MS patients was evaluated, using two-color flow cytometry before glucocorticosteroids and after 7 days from starting therapy. Significant increase of B cells, decrease of NK cells and monocytes producing IL-8 were observed after treatment. IL-8 is one of the cytokines responsible for blood-brain-barrier disruption and migration of immune cells to the central nervous system; in this aspect, explaining glucocorticosteroid effects during MS exacerbations.

Alternative methods of treatment used by multiple sclerosis patients in Poland.

BACKGROUND AND PURPOSE: Despite the great progress that has been made in medicine, treatment of many chronic diseases, including multiple sclerosis (MS), remains hardly effective. It has been reported that more and more MS patients worldwide are using alternative methods (AM). The aim of the present study is to evaluate this phenomenon in the Polish MS population. MATERIALS AND METHODS: To obtain more data about use of AM in Poland we distributed questionnaires among 210 MS patients treated in 3 hospitals in different regions of our country: Warsaw, Gdansk and Pila. RESULTS: 210 patients (136 females and 74 males) took part in the study. 68.5% (144 of 210 questioned) declared use of AM now or in the past. The most popular (56%) was Oenothera seed oil, followed by vitamins (49%), and the third most common was massage (34%). There were no significant demographic differences between users and non-users. Neither disease duration nor disability had any impact on the decision to use AM. CONCLUSIONS: The phenomenon of use of AM by MS patients is not fully understood, but it seems that limited access to so-called immunomodulators, e.g. interferon-b or glatiramer acetate, and limited long-term efficacy are reasons why patients seek other therapies.

[Comparison of the safety of the medicinal product in the European Union and the United States, tolcapone (Tasmar) -- COMT inhibitor as the analyzed example]. / Porównanie oceny bezpieczenstwa leku w Unii Europejskiej i Stanach Zjednoczonych na przykladzie inhibitora COMT -- tolkaponu (Tasmar).

The authors present regulatory procedures and pharmaco-vigilance systems obligatory in the European Union and the United States. The post-approval procedures in the European Union and the United States in case of serious drug reaction (urgent safety restriction) are discussed. Worldwide implementations of the post-approval safety procedure for the tolcapone case are analyzed. Some practical information concerning tolcapone and regulatory procedures obligatory in the European Union and the United States is included.

Cyclooxygenases mRNA and protein expression in striata in the experimental mouse model of Parkinson's disease induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine administration to mouse.

Cyclooxygenases (COX) are associated with complex alteration in many pathologies of the central nervous system (CNS). Increased expression of COX-2 has been shown in injured or degenerated neurons, thus suggesting that COX-2 may contribute to neuronal damage. In this study, we present the expression of COX-1 and COX-2 mRNA and protein in striatum following 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) administration to mice. MPTP causes an acute damage of dopaminergic neurons especially in the nigrostriatal dopaminergic system, thus diminishing dopamine (DA) content in striatum and decreasing the number of dopaminergic cells in the pars compacta of the substantia nigra (SN). C57Bl mice have received 60 mg/kg of MPTP introperitoneally. A group of mice received also rofecoxib 10 mg/kg from the 1st day following MPTP administration. Dopamine content in striatum (high-performance liquid chromatography-HPLC), mRNA expression of COX-1 and -2 (reverse transcriptase-polymerase chain reaction technique-RT-PCR), COX-1 and -2 protein content (immunoblotting) have been measured on day 1st, 3rd, 7th, 14th and 21st after the injury. We have found that COX-1 mRNA expression is not changed following MPTP administration, but COX-2 gene and protein expression in striatum increases from the 3rd to the 7th and 14th days, and diminishes on the 21st day. Production of prostaglandins is augmented only briefly after MPTP treatment and did not correlate with increased COX-2 mRNA and COX-2 protein production. Thus, the increase of COX-2 expression does not follow the acute stage of cell death but rather the recovery period after the injury. We also demonstrate that COX-2 activity inhibition by rofecoxib (10 mg/kg), which has been started 1 day after the injury, has not neuroprotective effect. Our study suggests that COX-2 does not contribute to neurons death following MPTP administration and that the inhibition of COX-2 activity is not beneficial to neurons injured by MPTP. However, COX-2 mRNA and protein expressions increase after MPTP injury; the role of these findings remains obscure.

Changes of percentages in immune cells phenotypes and cytokines production during two-year IFN-beta-1a treatment in multiple sclerosis patients.

OBJECTIVE: The aim of the study was to find out whether INF-beta-1a influences the immune profile of peripheral blood (PB) leukocytes in MS patients. METHOD: We have studied 20 patients with relapsing-remitting form of MS treated with INF-beta-1a using twocolor cytometry. We determined immune cells phenotypes and production of some cytokines: IL-4, IL-10, IL-12, IFN-gamma, before drug administration and after starting the treatment. RESULTS: In MS patients an increased percentage of CD14(+)CD86(+) cells and CD3(+)CD25(+) cells was noticed after 6, 9 and 12 months of INF-beta-1a therapy. Among cytokine-producing cells we noted an increased fraction of CD3(+)IL-4, CD14(+)IL-10 and CD14(+)IL-12 cells after 12 months, which decreased to the level observed before treatment after 24-month therapy. CONCLUSIONS: IFN-beta-1a treatment was associated with significant changes in immune response. This effect was mostly evident within the first year of treatment.

Dexamethasone protects against dopaminergic neurons damage in a mouse model of Parkinson's disease.

The pathological process of neurodegeneration, which is observed in Alzheimer's (AD) and Parkinson's (PD) diseases and that follows any insult to the central nervous system, is accompanied by an inflammatory reaction, which is believed to contribute to the pathogenesis of the diseases. In accordance to this, the anti-inflammatory agents are suggested to be effective in slowing or inhibiting the degenerative process. In this study, we investigated the influence of dexamethasone (DXM) on the nigrostriatal dopaminergic neurons damage following administration of 1-methyl-4-phenyl-1,2,3,6-tetrahydropiridine (MPTP). Mice C57BL received pre-treatment of the various doses of dexamethasone followed by MPTP administration (40 mg/kg). We found that dexamethasone 1 mg/kg diminished a dopamine content depletion in striatum by about 20%, when the doses of 0.1 mg/kg was ineffective and 10 mg/kg even aggravate the dopamine content decrease. In the second step of the experiment, we chose the effective doses, 1 mg/kg, and started the treatment before and 24 h after MPTP administration. We observed the same protection in both situations: less dopamine depletion and less decrease in the number of dopaminergic cells in the substantia nigra (SN). Dexamethasone also similarly decreased the inflammatory reaction (glial activation, lymphocytic infiltration) in the injured areas. Our study showed that dexamethasone may exert a neuroprotective effect towards neurons injured by MPTP, but only when used in a proper dose. The mechanism of dexamethasone protective properties may be an inhibition of inflammatory process; however, direct interactions with neurons are also possible.

Tryptase levels in patients after acute coronary syndromes: the potential new marker of an unstable plaque?

BACKGROUND: Mast cells (MCs) are multifunctional immune cells that produce a number of vasoactive or thromboactive mediators. Elevated numbers of human heart MCs are observed in the shoulder regions of coronary atherosclerotic plaques, suggesting that they play a role in plaque rupture. Cardiac MC degranulation after myocardial ischemia has been documented in animal models. Cardiac MCs are highly profibrinolytic cells and release tryptase, their specific protease, after ischemic events. HYPOTHESIS: Mast cell activation and release of tryptase may differentiate among patients with acute coronary syndromes (ACS), potentially determining the clinical course of ACS. Tryptase levels may indirectly reflect the fibrinolytic status of patients. METHODS: Mast cell activation after ACS was estimated in 10 controls and 52 patients by measuring the serum levels of tryptase in the acute phase, at 2 weeks, and at 3 months after the ACS episode. Total tryptase levels were determined by using the UniCAP system and analyzed with respect to the patients' clinical types of ACS on admission (ACS with persistent ST-segment elevation on electrocardiogram or with ST-segment depression). RESULTS: Significant differences in serum tryptase levels between the groups were found, with higher serum tryptase concentrations in the ST-segment depression group in the acute phase, and at follow-up. CONCLUSIONS: Serum tryptase concentration differences among patients with distinct types of ACS may indicate a more important role of human heart MCs in ACS with ST-segment depression pathogenesis. To our knowledge, this is the first report indicating that serum tryptase levels may differentiate patients with distinct types of ACS.

Post intoxicative therapeutic immunization with myelin oligodendrocyte glycoproteine (MOG 35-55) suppresses spontaneous regeneration of dopaminergic neurons injured with 1-methyl-4 phenyl-1,2,3,6-tetrahydropiridine (MPTP).

The pathological process of neurodegeneration is accompanied by an inflammatory reaction that is believed to contribute to the pathogenesis of neurodegenerative diseases. The aim of our study was to evaluate the influence of autoimmune reaction induced by post-traumatic vaccination with myelin self-antigen on spontaneous regeneration of dopaminergic neurons, injured with MPTP. C57BL mice were intoxicated with 40 mg/kg MPTP and seven days later immunized with MOG 35-55 peptide in CFA. On the 7th day following intoxication, the MPTP treated mice showed decrease of the dopamine level by 63% as compared to the control mice. However, starting from the 14th day following intoxication, a spectacular increase of dopamine content was observed. Immunization with MOG resulted in a statistically significant reduction of the increase in striatum as compared to non-immunized animals, and was lower by 23%, 17% and 15% on days 14, 28 and 50, respectively. Our results show suppressive influence of autoimmune reaction induced after injury on regeneration of dopamine cells intoxicated with MPTP.

Dynamics of expression of the mRNA for cytokines and inducible nitric synthase in a murine model of the Parkinson's disease.

The inflammatory reaction and oxidative stress has been linked with PD. Proinflammatory cytokines promote neurodegeneration or neuroprotection in different animal models. In addition, these cytokines have been reported to increase iNOS expression. With the RT-PCR method we evaluated mRNA levels for IL 1beta, IL6, TNF, IFNgamma, IL-10 and iNOS in the striatum of C57BL/6 mice after MPTP intoxication. The IL1beta mRNA expression rapidly increased and peaked at 6 h. The first increase of mRNA for TNFalpha and IFNgamma was noticed at 6-24 h and the second at the 7th day after MPTP intoxication. Two peaks of IL10 mRNA were seen, immediately (6 h) and at the 3 day post MPTP injection. The peak of mRNA level for IL6 was observed at the 7th day. Expression of mRNA for iNOS peaked at 24 h, started decreasing on the 3rd day, but was still present till the 14th day. Those findings suggest that cytokine network and iNOS may be involved in the development of immune changes accompanying degeneration of the nigrostriatal system.